Molecular phylogeny of the spiny-surfaced species of the dinoflagellate Prorocentrum with the description of P. thermophilum sp. nov. and P. criophilum sp. nov. (Prorocentrales, Dinophyceae).

Spiny-surfaced species of Prorocentrum form harmful algal blooms, and its taxonomic identity is obscure due to the size and shape variability. Molecular phylogenies reveal two major clades: one for P. cordatum with sequences mainly retrieved as P. minimum, and the other for P. shikokuense with sequences also retrieved as P. dentatum and P. donghaiense. Several closely related clades still need to be characterized. Here, we provide nuclear SSU and LSU rRNA genes, and nuclear ITS region (ITS1-5.8S gene-ITS2) sequences of the strain CCMP3122 isolated from Florida (initially named P. donghaiense) and strains Prorocentrum sp. RCC6871-2 from the Ross Sea, Antarctica. We describe Prorocentrum thermophilum sp. nov. based on the strain CCMP3122, a species also distributed in the open waters of the Gulf of Mexico, New Zealand, and the Arabian Gulf; and Prorocentrum criophilum sp. nov. based on the strain RCC6872, which is distributed in the Antarctic Ocean and Arctic Sea. Prorocentrum thermophilum is roundish (~14 µm long, ~12 µm wide), with an inconspicuous anterior spine-like prolongation under light microscopy, valves with tiny, short knobs (5-7 per µm2 ), and several (<7) large trichocyst pores (~0.3 µm) in the right valve, as well as smaller pores (~0.15 µm). Prorocentrum criophilum is round in valve view (~11 µm long, 10 µm wide) and asymmetrically roundish in lateral view, the periflagellar area was not discernible under light microscopy, valves with very tiny, short knobs (6-10 per µm2 ), and at least 12 large pores in the right valve. Other potentially undescribed species of spiny-surfaced Prorocentrum are discussed.

Playing hide and seek: Distribution with depth of potentially harmful epibenthic dinoflagellates of Southern El Hierro Island, Canary Islands (NE Atlantic).

The study of epibenthic assemblages of harmful dinoflagellates (BHABs) is commonly conducted in shallow infralittoral zones (0 - 5 m) and are seldom investigated at deeper waters. In this study, the distribution with depth of five BHAB genera (Gambierdiscus, Ostreopsis, Prorocentrum, Coolia and Amphidinium) was investigated in the south of El Hierro island (Canary Islands, Spain). Sampling involved the use of a standardized artificial substrate deployed at three depth levels (5, 10 and 20 m) that were visited at three different times throughout one year. The influence of three depth-correlated abiotic parameters, i.e. light, water motion and water temperature, on the vertical and seasonal distribution of the BHAB assemblage was also assessed. Two vertical distribution patterns were observed consistently through time: cell abundances of Ostreopsis and Coolia decreased from 5 to 20 m while those of Gambierdiscus, Prorocentrum and Amphidinium showed the reverse pattern, although significant differences were only observed between 5 and 10 - 20 m depth. In April, two members of the latter group, Gambierdiscus and Amphidinium, were even absent at 5 m depth. The recorded environmental parameters explained a high percentage of the observed distribution. In particular, model selection statistical approaches indicated that water motion was the most significant parameter. An analysis of Gambierdiscus at species level revealed the co-occurrence of four species in the study area: G. australes, G. belizeanus, G. caribaeus and G. excentricus. The species G. excentricus, reported here for the first time in El Hierro, showed a more restricted vertical and seasonal distribution than the other species, which may explain not being detected in previous studies in the area. The results obtained in this study highlight the importance of considering a wider depth range and different seasons of the year when investigating the ecology of BHABs and assessing their risk and impacts on human health and the environment. Only then, efficient monitoring programs will be implemented in the Canary Islands and globally in areas affected by these events.

Using artificial substrates to quantify Gambierdiscus and other toxic benthic dinoflagellates for monitoring purposes.

Collecting methods generally used to determine cell abundances of toxic benthic dinoflagellates (BHAB) use cells dislodged from either macrophytes or artificial substrates. This article compares the advantages of the macrophyte and artificial substrate methods and discusses which method is more appropriate for use in monitoring programs that focus on toxic BHAB species identification and quantification. The concept of benthic dinoflagellate "preference" for specific macrophytes was also reviewed. Examination of data from 75 field studies showed macrophytes with higher surface area per unit biomass harbored higher concentrations of Gambierdiscus cells. There was no definitive evidence that cells were actively selecting one macrophyte over another. This observation supports the use of artificial substrates (AS) as a means of assessing cell abundances in complex habitats because cell counts are normalized to a standardized surface area, not macrophyte biomass. The artificial substrate method represents the most robust approach, currently available, for collecting toxic, benthic dinoflagellates for a cell-based early warning system.

Haslea silbo, A Novel Cosmopolitan Species of Blue Diatoms.

Specimens of a new species of blue diatoms from the genus Haslea Simonsen were discovered in geographically distant sampling sites, first in the Canary Archipelago, then North Carolina, Gulf of Naples, the Croatian South Adriatic Sea, and Turkish coast of the Eastern Mediterranean Sea. An exhaustive characterization of these specimens, using a combined morphological and genomic approach led to the conclusion that they belong to a single new to science cosmopolitan species, Haslea silbo sp. nov. A preliminary characterization of its blue pigment shows similarities to marennine produced by Haslea ostrearia, as evidenced by UV-visible spectrophotometry and Raman spectrometry. Life cycle stages including auxosporulation were also observed, providing data on the cardinal points of this species. For the two most geographically distant populations (North Carolina and East Mediterranean), complete mitochondrial and plastid genomes were sequenced. The mitogenomes of both strains share a rare atp6 pseudogene, but the number, nature, and positions of the group II introns inside its cox1 gene differ between the two populations. There are also two pairs of genes fused in single ORFs. The plastid genomes are characterized by large regions of recombination with plasmid DNA, which are in both cases located between the ycf35 and psbA genes, but whose content differs between the strains. The two sequenced strains hosts three plasmids coding for putative serine recombinase protein whose sequences are compared, and four out of six of these plasmids were highly conserved.

Benthic dinoflagellates: Testing the reliability of the artificial substrate method in the Macaronesian region.

The suitability of the 'artificial substrate' method, i.e. standardized surfaces of fiberglass screens, for the quantification of four benthic harmful algal bloom (BHAB) dinoflagellates (Gambierdiscus, Ostreopsis, Prorocentrum and Coolia) was tested relative to estimates from natural macroalgal substrates. Sampling took place in a variety of intertidal and subtidal coastal habitats under different water motion conditions, at depths from 1 to 7 m, in two archipelagos of the Macaronesia region: The Canary Islands and Cape Verde. An immersion time of 24 h was sufficient to adequately estimate dinoflagellate abundances. Seven replicates were established as the ideal replication level, considering both reproducibility and sampling effort. In most cases, cell abundances of the four dinoflagellate genera showed lower variability on artificial substrates than on macroalgae, leading to more reliable estimates of abundances. The ratio of mean cell abundances on artificial substrates to mean cell abundances on macroalgae highly varied among sampling sites for each genus. This was especially true for Ostreopsis and Coolia. Thus, given the potentially harmful nature of benthic dinoflagellates, the transformation of abundances expressed as cells g-1 of macroalgae to abundances expressed as cells cm-2 is risky, and it should not be attempted in monitoring and management programs of harmful microalgae. In summary, results of this study support the use of artificial substrates in monitoring programs of BHAB dinoflagellates, while the risks of using macroalgae are stressed.